目的 紫芝Ganoderma sinense次生代谢产物含量较低，筛选一种可以显著提高紫芝次生代谢物产量的方法。方法 基于紫芝基因组数据库筛选出编码芳樟醇合酶的基因GsSTS50，通过改善融合酶、共表达基因和引入异源甲羟戊酸（mevalonate pathway，MVA）代谢途径等方法使GsSTS50基因在大肠杆菌中高效表达，利用顶空固相微萃取-气相色谱-质谱联用（HS-SPME-GC-MS）检测大肠杆菌培养物中芳樟醇的含量。结果 优化融合酶的策略，使大肠杆菌中芳樟醇产量提高了约4.9倍；通过引入异源MVA途径，改造后菌株芳樟醇产量比初始培养提高了约49倍；将多个基因共表达的同时引入异源途径，优化菌株芳樟醇产量比原始菌株提高了42倍。结论 经过改造的重组菌株生产产物的量均有显著提高，为后续灵芝等担子菌中萜类化合物以及其他萜类物质产量的提升提供了参考。

Objective The low content of secondary metabolites of Ganoderma sinense has brought some difficulties to the development of monomers, so it is necessary to screen a method that can significantly increase the yield of secondary metabolites of G. sinense. Methods The GsSTS50 gene encoding linalool synthetase was selected based on the G. sinense gene library. The GsSTS50 gene was overexpressed in Escherichia coli by co-expression and introduction of heterologous mevalonate (MVA) pathway. The linalool content in E. coli was determined by HS-SPME-GC-MS. Results The yield of linalool and other terpenoids in E. coli was increased by about 4.9 times by optimizing the strategy of fusion enzyme. By introducing heterologous MVA pathway, the production of linalool was 49 times higher than that of the original culture. The co-expression of multiple genes at the same time into the heterologous pathway, the yield of the optimized strain was 42 times higher than that of the original strain. Conclusion The results showed that the yield of recombinant strains was significantly increased. This study provides a reference for further research on terpenoids and other terpenoids from G. sinense.

R286.2

国家自然科学基金资助项目（81603221）